Proliferation and harvesting of secondary protocorms as a novel means for improving propagation of terrestrial orchids

2014 ◽  
Vol 62 (7) ◽  
pp. 614 ◽  
Author(s):  
Betty Mauliya Bustam ◽  
Kingsley Dixon ◽  
Eric Bunn
Keyword(s):  
Early Stage ◽  
Naphthaleneacetic Acid ◽  
Novel Approach ◽  
Stage 4 ◽  
Terrestrial Orchids ◽  
Half Strength ◽  
Conservation Concern ◽  
Basal Salts ◽  
Proliferation In Vitro

This study investigated optimisation of media and primary-protocorm development stages to enhance secondary-protocorm production as a novel means for propagation of terrestrial orchids, including taxa of conservation concern. Seeds of Caladenia latifolia were germinated asymbiotically on ½-strength Murashige and Skoog (MS) medium fortified with 5% (v/v) coconut water. Resulting protocorms at 3, 5 and 7 weeks of growth were subcultured to protocorm-proliferation media treatments consisting of ½-strength MS basal-salts medium with 6-benzylaminopurine (BA) and α- naphthaleneacetic acid (NAA) singly or in combination. Conversion of seeds to primary protocorms was high (87–92%). The highest percentage of secondary-protocorm proliferation was 40.1%, using 5-week-old protocorms (early Stage 4 of protocorm development) as explants and cultured on ½-strength MS with a combination of 5 µM NAA + 2 µM BA. Half-strength MS media containing a single plant-growth regulator (BA or NAA) were substantially less effective (<10% protocorm proliferation). The present study has provided a novel approach to sequential protocorm production that will be of value particularly for threatened orchids with limited seed availability. Protocorm proliferation in vitro enables a renewable supply of protocorms with which to conduct propagation, cryostorage and pilot restoration programs.

scholarly journals Micropropagation of White Flowering Eastern Redbud (Cercis canadensis var. alba L.)

1990 ◽  
Vol 8 (4) ◽  
pp. 177-179
Author(s):  
S. Yusnita ◽  
R. L. Geneve ◽  
S. T. Kester
Keyword(s):  
Root System ◽  
Day Treatment ◽  
Shoot Multiplication ◽  
Naphthaleneacetic Acid ◽  
Indolebutyric Acid ◽  
Cercis Canadensis ◽  
Half Strength ◽  
Eastern Redbud ◽  
Being Moved

Abstract A white flowering Eastern redbud (Cercis canadensis var. alba L.) has been successfully micropropagated. Two node explants collected from the initial flush of spring growth were cultured on woody plant medium (WPM). Increased shoot multiplication occurred at 10,15 and 20 μM (2.3, 3.4 and 4.5 ppm) benzyladenine (BA). Microshoots were rooted in vitro on half strength WPM with a 15-day treatment of 100 and 300 μM (18.6 and 55.9 ppm) α-naphthaleneacetic acid (NAA) or 100 and 300 μM (20.3 and 60.9 ppm) indolebutyric acid (IBA) prior to being moved to full strength WPM without growth regulators. Percentage rooting and the mean number of roots per cutting were comparable between NAA and IBA treated microcuttings, however, the subsequent root morphology differed between the two treatments. NAA treated plants developed a coarse, unbranched root system, while IBA treated cuttings developed a more desireable fine, branched root system. Rooted microshoots were successfully acclimated to greenhouse conditions.

scholarly journals Physiological Response of In Vitro Cultured MAGNOLIASP. to Nutrient Medium Composition

2014 ◽  
Vol 22 (1) ◽  
pp. 49-61 ◽  
Author(s):  
Rossen S. Sokolov ◽  
Bistra Y. Atanassova ◽  
Elena T. Iakimova
Keyword(s):  
Nutrient Medium ◽  
Culture Conditions ◽  
Medium Composition ◽  
Naphthaleneacetic Acid ◽  
Axillary Shoots ◽  
Regeneration Rate ◽  
Primary Explants ◽  
High Regeneration ◽  
Basal Salts

AbstractThe objective of this study was to assess the regeneration response of in vitro cultured Magnolia × soulangeana ‘Alexandrina’ and Magnolia liliiflora ‘Nigra’ to nutrient medium composition. In the primary culture (initiated from dormant axillary buds) combinations of Murashige and Skoog (MS) basal salts with 6-benzylaminopurine and α-naphthaleneacetic acid were tested. The primary explants of cv. ‘Alexandrina’ expressed higher regeneration rate than cv. ‘Nigra’. For both species, the regen eration was most strongly potentiated at addition of 0.25 mg dm−3 of the cytokinin alone. The auxin exerted undesir–able effects. Several basal salts media were applied in proliferation stage and their physiological effects were evaluated in reference to traditionally used MS. At culturing on Chée & Pool C2d Vitis Medium (VM) that is for the first time introduced to magnolia and on MS, M. liliiflora formed more but less elongated shoots than M. soulangeana. However, on VM, substantial increase (25-30%) of the number of axillary shoots and leaves, shoot length and fresh and dry weights over MS was established for both species. This suggested VM as promising composition of nutrients in multiplication stage. Microshoots obtained on MS, VM, Rugini Olive Medium and DKW Juglans Medium were successfully rooted in vitro and subsequently established ex vitro. The findings expand the information on magnolia response to culture conditions and contribute to elaboration of innovative elements of protocols for establishing tissue cultures with high regeneration capacity.

scholarly journals Conservation-driven Propagation of an Epiphytic Orchid (Epidendrum nocturnum) with a Mycorrhizal Fungus

HortScience ◽  
2007 ◽  
Vol 42 (1) ◽  
pp. 135-139 ◽  
Author(s):  
Lawrence W. Zettler ◽  
Sarah B. Poulter ◽  
Kris I. McDonald ◽  
Scott L. Stewart
Keyword(s):  
Mycorrhizal Fungus ◽  
Leaf Length ◽  
Ex Vitro ◽  
Commercial Fertilizer ◽  
Epiphytic Orchid ◽  
Seed Sources ◽  
Terrestrial Orchids ◽  
Half Strength ◽  
Symbiotic Seed Germination

Seeds of an endangered epiphytic orchid from Florida (Epidendrum nocturnum Jacquin) germinated in vitro with a mycorrhizal fungus [Epulorhiza repens (Bernard) Moore] using a technique normally applied to terrestrial orchids (symbiotic seed germination). Seeds from two sources (Fakahatchee Strand, Fla. Panther NWR) were sown on either modified oats medium (MOM) or standard oat medium (SOM) and inoculated with the fungus. Significant differences in germination were detected between the two seed sources. MOM had a significant effect on mean leaf length during incubation in vitro (F (1278) = 23.81, P > 0.000), but media had no significant effect on leaf number. After 48 days in vitro, all leaf-bearing seedlings were exposed to light and then transferred to greenhouse conditions ex vitro on sterile Sphagnum moss with or without half-strength Miracle-Gro (Scotts, Port Washington, N.Y.) commercial fertilizer. After 163 days ex vitro, seedlings on Sphagnum without Miracle-Gro displayed highest survivorship (>90%), whereas Miracle-Gro-exposed seedlings from standard oat agar experienced low (44%) survivorship. Healthy seedlings with a mycotrophic capability were obtained 1 year after sowing. A total of 43 seedlings were subsequently reintroduced into the Florida Panther NWR in Nov. 2005, 16 months after sowing. The symbiotic technique may, therefore, have practical merit for conservation of E. nocturnum and other epiphytic orchids threatened with extinction.

scholarly journals Pomegranate Micropropagation, Callogenesis and Genetic Integrity Assessment Using Simple Sequence Repeat Markers

2018 ◽  
Vol 11 (1) ◽  
pp. 237
Author(s):  
Tebogo Stimela ◽  
Remmy W. Kasili ◽  
Edward G. Mamati
Keyword(s):  
Acetic Acid ◽  
Simple Sequence Repeat ◽  
Naphthaleneacetic Acid ◽  
Genetic Integrity ◽  
Sequence Repeat ◽  
Simple Sequence Repeat Markers ◽  
Half Strength ◽  
Indole 3 Acetic Acid ◽  
Simple Sequence

In recent years, the awareness of pomegranate health benefits has grown exponentially; nonetheless the existing propagation methods remain a challenge to supply adequate suitable planting materials needed for commercial production. Micropropagation can lead to mass production of plantlets and callus-mediated in vitro regeneration can open avenues for the use of genetic engineering to improve this crop. The aim of this study was to evaluate appropriate conditions for pomegranate micropropagation, callogenesis and use Simple Sequence Repeat markers to screen for somaclonal variation. Cytokinins (Benzylaminopurine, Kinetin and Thiadiazol-5ylurea) were tested for shoot induction from nodal explants while auxins (1-Naphthaleneacetic acid, Indole-3-butyric acid and Indole-3-acetic acid) were tested for root induction of in vitro regenerated shoots. 1-Naphthaleneacetic acid combined with Benzylaminopurine was assessed for their ability to induce callus from cotyledon and leaf explants. Genetic integrity between mother plant, callus and in vitro regenerated shoots were assessed using eight Simple Sequence Repeat markers. Maximum number of shoots and leaves were obtained on full strength Murashige and Skoog media with 6.9 &micro;M kinetin. The highest number of roots was achieved on half strength Murashige and Skoog media with 4.9 &micro;M Indole-3-butyric acid and the longest root was got on half strength Murashige and Skoog media with 5.3 &micro;M Indole-3-acetic acid. Leaves and cotyledons demonstrated to be potential explants for callus formation at all hormonal combination levels tested. Eight out of 13 amplified alleles were polymorphic. A wider genetic variation was found with similarity coefficient range of 0.46-0.92. More somaclonal variation was in regenerated shoots compared to callus.

scholarly journals Explant Initiation Date and BA Concentration Influence Shoot Proliferation in vitro of Two Liatris Interspecific Hybrids

HortScience ◽  
2004 ◽  
Vol 39 (5) ◽  
pp. 1098-1100 ◽  
Author(s):  
James R. Ault
Keyword(s):  
Shoot Growth ◽  
Shoot Proliferation ◽  
Shoot Formation ◽  
Axillary Shoot ◽  
Stem Explants ◽  
Hybrid Plants ◽  
Flower Stem ◽  
Basal Salts ◽  
Proliferation In Vitro

Shoot proliferation cultures were established in vitro using flower-stem explants from two different interspecific hybrid plants of Liatris. Explants taken on two dates from field-grown plants were successfully established and axillary shoot growth promoted on a medium consisting of Murashige and Skoog basal salts and vitamins with 30 g·L-1 sucrose, 1.0 μm BA, and 7.0 g·L-1 agar, with a medium pH = 5.7. Initial explant contamination rates were significantly higher among explants collected later in the growing season. Addition of BA (1.0, 2.0, 4.0, 8.0, or 16.0 μm) improved shoot formation compared to the control for both plants. Proliferation rates differed between the dates of establishment, the plants, and the BA treatments. Shoots rooted readily in medium without PGRs or with 1.0, 2.0, 4.0, or 8.0 μm K-IBA. Overall rooting was 88%. About 90% of the plants rooted in the presence of 1.0 μm K-IBA were successfully established in the greenhouse. Chemical names used: 6-benzyl adenine (BA); potassium salt of indole-3-butyric acid (K-IBA).

In Vitro Reproductive Development of a Diploid Wild Species, Arachis duranensis1

1994 ◽  
Vol 21 (2) ◽  
pp. 139-143
Author(s):  
Q. L. Feng ◽  
H. E. Pattee ◽  
H. T. Stalker
Keyword(s):  
Growth Regulators ◽  
Early Stage ◽  
Basal Medium ◽  
Reproductive Development ◽  
Naphthaleneacetic Acid ◽  
Embryo Abortion ◽  
Culture Techniques ◽  
Combination Treatments ◽  
Four Levels

Abstract Embryo abortion at an early stage of reproductive development is a major impediment for introgressing germplasm from wild to cultivated species of Arachis by interspecific hybridization. Ovule and embryo culture techniques have been used to rescue aborting hybrid embryos, but increased efficiency and recovery of very young tissues are still needed. The objective of this study was to induce growth and differentiation of A. duranensis proembryos. Seven-, 10-, and 14-d-old peg tips were cultured on a modified basal medium containing MS and B5 media combinations with 16 combination treatments using three growth regulators—1-naphthaleneacetic acid, gibberellic acid, and 6-benzylaminopurine—each at four levels. The results showed that seeds could be obtained in vitro by peg tip culture of four- to 16-celled proembryos. The favorable concentration ranges of growth regulators for pod formation and embryo development were 0.5-2.0 mg/L NAA, 0.05-0.5 mg/L GA3, and 0.05-0.2 mg/L 6-BAP. Over all three selected ages of pegs, the three best combinations of growth regulators resulted in 4.8, 4.7, and 3.5% pod formation, respectively.

In vitro propagation of Epacris impressa (Epacridaceae) and the effects of clonal material

2000 ◽  
Vol 48 (2) ◽  
pp. 215 ◽  
Author(s):  
J. Anthony ◽  
C. B. McLean ◽  
A. C. Lawrie
Keyword(s):  
In Vitro Propagation ◽  
Root Formation ◽  
Woody Plant ◽  
Multiple Shooting ◽  
Naphthaleneacetic Acid ◽  
Root Induction ◽  
Main Determinant ◽  
Woody Plant Medium ◽  
Half Strength

A system of micropropagation has been developed for Epacris impressa Labill. (pink heath) (Epacridaceae), the floral emblem of Victoria, Australia. Only explants from glasshouse-grown plants treated with 1.2 g L–1 mancozeb were established successfully in vitro. Shoot material was very sensitive to surface-sterilisation, with 0.5% NaOCl for 5 min being optimal. Multiple shooting was induced optimally on Woody Plant Medium (WPM, Lloyd and McCown 1980) with 12–25 µM of the cytokinin 2iP (6-(γ,γ-dimethylallylamino) purine). Inclusion of the auxin IBA (indole-3-butyric acid) induced callus and reduced shooting. Rooting in vitro was greatest (up to 40%) with half-strength WPM and 16 µM IBA. Clones from individual plants varied in multiple shooting response to 2iP (0–49 µM) and root induction response to auxins (IBA and NAA (α-naphthaleneacetic acid), 0–43 µM). These results suggest that explant materials are the main determinant of success in in vitro propagation and that they require individual optimisation of treatments to maximise shoot and root formation.

scholarly journals In vitro Propagation of Solanecio biafrae and Determination of Genetic Stability of Plantlets Using RAPD and ISSR Markers

2016 ◽  
Vol 24 (1) ◽  
pp. 29-36 ◽  
Author(s):  
Jelili Opabode ◽  
Oluyemisi Akinyemiju
Keyword(s):  
Random Amplified Polymorphic Dna ◽  
Issr Markers ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Germplasm Exchange ◽  
Indole Butyric Acid ◽  
Half Strength ◽  
Micropropagated Plants

Abstract An efficient and reproducible micropropagation protocol of Solanecio biafrae (Oliv. & Hiern) C. Jeffrey has been developed from nodal stem segments. Shoot development was obtained on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP) alone and in combination with zeatin and 1-naphthaleneacetic acid (NAA). Elongated shoots were rooted in the presence of zeatin or 3-indole-butyric acid (IBA) alone or in combinations. The highest number of explants forming shoots (100%) as well as the highest number of shoots per explant (3.4) and the longest shoots (22 mm) were recorded on medium containing 4.0 mg·dm−3 BAP, 2.0 mg·dm−3 NAA, and 1.0 mg·dm−3 zeatin. About 76% of shoots formed roots on half-strength MS medium free of plant growth regulators. The best root formation (approximately 88%) was recorded on the medium containing 1.0-1.5 mg·dm−3 IBA. The micropropagated shoots with well-developed roots were efficiently acclimatized under greenhouse conditions. The random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) amplification products were monomorphic in micropropagated plants and similar to those of mother plant showing their genetic uniformity. This is the first report of micropropagation of S. biafrae, which will facilitate in vitro mass propagation, conservation, and germplasm exchange of this endangered African vegetable.

scholarly journals ENHANCED REGENERATION OF SHOOTS FROM PEACH CELLS INFECTED WITH A SHOOTY MUTANT STRAIN OF AGROBACTERIUM.

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1150d-1150
Author(s):  
A. Smigocki ◽  
F. Hammerschlag
Keyword(s):  
Prunus Persica ◽  
Northern Analysis ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Gene Transcripts ◽  
Half Strength ◽  
Low Levels ◽  
2,4 Dichlorophenoxyacetic Acid

Immature `Redhaven' peach (Prunus persica L. Batsch) embryos were infected with Agrobacterium tumefaciens strain tms328::Tn5 carrying the functional cytokinin gene. Shoots were regenerated from callus grown on MS medium without added phytohormones and subsequently rooted on half-strength MS medium with 2.8 -naphthaleneacetic acid. These plants exhibited an increased frequency of branching in vitro. Low levels of cytokinin gene transcripts were detected in these cells by Northern analysis, and using an ELISA assay, the cytokinins zeatin and zeatinriboside were determined to be on the average 30-fold higher. From these results, the expression of the cytokinin gene appears to promote growth of cells in the absence of phytohormones thus serving as a marker for transformation and a promoter of morphogenesis without a 2,4-dichlorophenoxyacetic acid inductive step.

scholarly journals Studies on Seed Germination and Micropropagation of Clinopodium nepeta: A Medicinal and Aromatic Plant

HortScience ◽  
2019 ◽  
Vol 54 (9) ◽  
pp. 1558-1564 ◽  
Author(s):  
Georgia Vlachou ◽  
Maria Papafotiou ◽  
Konstantinos F. Bertsouklis
Keyword(s):  
Shoot Multiplication ◽  
Adult Plant ◽  
Naphthaleneacetic Acid ◽  
Similar Response ◽  
Ms Medium ◽  
Rooting Medium ◽  
Shoot Number ◽  
Half Strength ◽  
Mediterranean Plant

Seed ecophysiology and micropropagation of Clinopodium nepeta, an aromatic Mediterranean plant with pharmaceutical and horticultural uses was investigated. The optimum germination temperature of seeds stored at room temperature for 0, 6, or 12 months was 15 to 20 °C (100% germination completed in 10 to14 days) and cardinal temperatures were defined at 10 and 30 °C (80% to 82% and 62% to 76% germination, respectively). Six or 12 months of storage did not seem to affect germination, although 12-month-old seeds germinated at higher percentage and completed germination earlier at 15 °C than at 20 °C. Concerning micropropagation, shoot multiplication at subcultures of both adult plant- and seedling-origin nodal explants was tested on Murashige and Skoog (MS) medium supplemented with various cytokinin types, i.e., zeatin (ZEA), 6-benzyladenine (BA), kinetin (KIN), and 6-γ-γ-(dimethylallylamino)-purine (2IP), at various concentrations from 0.0 to 8.0 mg·L−1. Both explant types presented a rather similar response during in vitro culture. Increasing concentration of all cytokinin types resulted in an increase in shoot number per responding explant (1.1–5.3) and in most cases a decrease in shoot length (0.6–3.4 cm). Increasing cytokinin concentration induced hyperhydricity to a number of shoots (0.1–6.5) per explant, mostly when ZEA and BA were used. Supplementing the MS medium with 8.0 mg·L−1 BA combined with 0.1 mg·L−1 1-naphthaleneacetic acid (NAA) led to almost elimination of hyperhydricity and very satisfactory shoot production (80%/88% explant response and 6.5/7.5 shoot number per responding explant for seedling- / adult-origin explants, respectively). Alternatively, increasing the agar concentration to 12.0 g·L−1 and supplementing the medium with 8.0 mg·L−1 BA only, resulted in the same effect on eliminating hyperhydricity, such as the addition of NAA, and in the best shoot multiplication response achieved in this study (100% explant response, 9.4/9.9 shoots per explant for seedling-/adult-origin explants, respectively). Microshoots rooted abundantly (92% to 100%) on half-strength MS medium, either Hf or supplemented with 0.5 mg·L−1 to 4.0 mg·L−1 indole-3-butyric acid (IBA). The addition of IBA to the rooting medium, regardless of its concentration, affected only the root length by increasing it 2- to 3-fold. Microshoot clusters produced on multiplication media rooted at 96% when cultured on Hf half-strength MS medium. Rooted microshoots and shoot clusters survived at 80% to 100%, respectively, after ex vitro acclimatization in peat:perlite 1:1 (v/v).

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